实时定量RT-PCR对乙型肝炎病毒全长RNA(fRNA)的定量检测

目的:建立一种简便的定量检测慢性乙型肝炎患者血清中终止于聚腺苷酸化位点的乙型肝炎病毒全长RNA(f RNA)的方法。方法:选取53例未治疗的乙型肝炎患者及22例HBs Ag阴性的健康者为研究对象,使用锚定oligo-d T的引物对其血清中f RNA进行实时定量反转录PCR检测,统计分析其与HBV DNA、HBcr Ag和HBe Ag的相关性。结果:对f RNA进行实时荧光定量RT-PCR检测的下线为2.3 log copies/ml,标准曲线的相关系数为0.99(P0.0001)。53例乙型肝炎患者中,29例(54.7%)可以检测到f RNA,22例正常对照中没有检测到f RNA。27例HBe Ag阳性和/或高水平HBV DNA的患者全部检测到f RNA,26例HBe Ag阴性并且低水平HBV DNA的乙型肝炎患者中有2例(7.7%)检测到f RNA(P0.0001)。HBe Ag阳性患者血清中f RNA水平高于HBe Ag阴性患者(5.0±0.3 vs.2.9±0.4 log copies/ml,P0.001)。f RNA与HBV DNA/HBcr Ag具有显著相关性(r=0.905、0.881,P0.0001)。Hayashi's定量分析法I显示f RNA与HBV DNA相关性强于其与HBcr Ag的相关性。结论:与HBV DNA和HBe Ag一样,f RNA可作为常规检测判断HBV的复制水平并指导用药。     

乙型肝炎病毒B,C基因型引起的慢性乙型肝炎患者特异性、非特异性细胞免疫比较

对138例CHB患者进行HBV基因分型,检测非特异性CTL,HBV特异性CTL,肝功能,HBVDNA,HBVM(HBsAg,抗-HBs,HBeAg,抗-HBe,抗-HBc).结果显示,C基因型感染者,非特异性CTL((19.91±6.01)%)高于B基因型感染者((16.12±3.12)%),t=3.05,P<0.01,HBVDNA水平((6.25±0.81)log10拷贝/mL)高于B基因型感染者((5.02±0.61)log10拷贝/mL),t=6.03,P<0.01,HBeAg阳性46例(74.19%),高于B基因型感染者(39例,占52%),χ2=7.09,P<0.01,丙氨酸氨基转移酶(ATL)、血清总胆红素(TBIL)水平(分别为(507.15±312.17)IU/L和(46.10±40.18)μmol/L)高于B基因型感染者(分别为(290.05±215.12)IU/L和(29.12±17.12)μmol/L),t=3.76,2.29,P<0.01,P<0.05.C基因型感染者的HBV特异性CTL((0.21±0.05)%),低于B基因型感染者((0.39±0.12)%),t=5.61,P<0....     

HBsAg定量检测在慢性乙型肝炎自然病程中的意义

目的 研究慢性乙型肝炎病毒(HBV)感染自然病程中HBsAg水平变化情况以及和HBV DNA的关系.方法 将183名HBV感染者按感染的不同阶段分为免疫耐受期(IT)42例,免疫清除期(IC)46例,非/低水平复制期(LR)57例,和再活动期(HBeAg阴性肝炎组ENH)38例,回顾性分析每个阶段血清中HBsAg和HBV DNA水平以及两者之间的关系.结果 HBsAg含量分别为IT期最高[(3.81±0.46) Ig IU/mL],其次为IC期[(3.41±0.77) Ig IU/mL],最低是LR期[(2.87±0.61)Ig0.61IU/mL],每期之间HBsAg含量差异有统计学意义(P＜0.05),ENH组HBsAg含量又有所升高[(3.31 ±0.78)Ig IU/mL].IT期HBsAg与HBV DNA无相关(r=0.231,P＞0.05),IC期HBsAg与HBV DNA呈正相关(r=0.612,P＜0.05),ENH期两者无相关(r=0.189,P＞0.05);不同时期HBsAg含量与ALT均无相关性.结论 HBV感染的不同阶段HBsAg水平有差异,IT期最高,进入IC期后开始下降,在LR期降至最低,ENH期再度升高,HBsAg与HBV DNA仅在IC期有相关性,血清HBsAg的表达与不同的免疫背景有关.     

基线血清外泌体miR-155-5p联合乙型肝炎病毒DNA定量实验预测聚乙二醇干扰素治疗乙型肝炎e抗原阳性慢性乙型肝炎的疗效

探讨聚乙二醇干扰素(peginterferon, Peg-IFN)治疗乙型肝炎e抗原(hepatitis B e antigen, HBeAg)阳性慢性乙型肝炎(chronic hepatitis B, CHB)患者48周后,基线血清外泌体miR-155-5p表达水平联合乙型肝炎病毒DNA定量实验在预测HBeAg血清学转换中的价值。回顾性分析2016年6月至2019年6月在本中心初次接受抗病毒治疗的HBeAg阳性CHB患者88例。根据Peg-IFN治疗48周后是否发生HBeAg血清学转换,将患者分为治疗应答组和无应答组。采用多因素logistic回归探讨预测Peg-IFN治疗应答的因子,并应用受试者工作特征(receiver operating characteristic, ROC)曲线下面积评估预测效能。结果发现基线血清外泌体miR-155-5p(OR＝2.193,95% CI 1.315～3.655,P＝0.003)和HBV DNA(OR＝0.398,95% CI 0.163～0.976,P＝0.036)是Peg-IFN治疗效果的独立预测因子。基线血清外泌体miR-155-5p和HBV DNA的截断值分别取2.3和7.2 log₁₀IU/mL时,相应的ROC曲线下面积分别为0.788(95% CI 0.682～0.893)和0.704(95% CI 0.577～0.824)。基线血清外泌体miR-155-5p表达水平≥2.3且HBV DNA定量≤7.2 log₁₀IU/mL的患者,Peg-IFN治疗48周后其HBeAg血清学转换率最高,为66.67% (10/15);而基线血清外泌体miR-155-5p表达水平<2.3且HBV DNA定量>7.2 log₁₀IU/mL的患者,Peg-IFN 治疗48周后其HBeAg血清学转换率最低,仅为3.03%(1/33)。这些结果表明,基线血清外泌体miR-155-5p表达水平联合HBV DNA定量实验可以作为Peg-IFN 治疗HBeAg阳性CHB疗效的预测因子,对于优化CHB的抗病毒治疗有积极作用。     

慢性乙型肝炎治疗的适应症和治疗方案的最新进展

目前,对于治疗一些慢性乙型肝炎病毒(HBV)感染边缘病例的最佳方法尚存在争议.血清HBV DNA和转氨酶水平、炎性坏死的程度和肝硬变程度决定着治疗的方案.所有转氨酶升高(＞正常上限值2倍)和血清HBVDNA＞20000 IU/mL的患者都需进行治疗.肝脏活检对于制定转氨酶轻度升高和血清HBV DNA＜20000 IU/mL的病例的治疗决策非常重要.慢性HBV患者如未接受治疗则需长期随访.现有7种药物获批用于治疗慢性乙型肝炎:普通和聚乙二醇IFNα、拉米夫定、阿德福韦、恩替卡韦、替比夫定和替诺福韦.应用聚乙二醇IFNα 1年疗程后持续治疗后效应使HbeAg阳性慢性乙型肝炎患者缓解率为30%-32%.口服抗病毒药物适用于大部分患者,并被用于长期治疗.然而,病毒耐药是长期口服抗病毒药物的主要缺点.     

HBV定量检测对拉米夫定停药的指导意义

目的:探讨核苷类似物(NAs)拉米夫定(LAM)用药时间与乙型肝炎(CHB)患者体内HBV病毒血清HBV-DNA载量的关系。方法:选取214例HBeAg阳性患者,在LAM(100 mg/d)的36个月用药治疗前使用实时荧光定量PCR进行血清HBV-DNA载量进行测定,并使用特异性引物鉴定HBV病毒DNA保守位点YMDD的抗药性突变以及病毒株的变化情况。结果:(1)214例患者均对LAM产生应答,在治疗12-18个月后血清HBV-DNA载量稳定在较低水平。(2)病毒株在NAs治疗前已经出现耐药性突变(18%),用药加速了HBV病毒的耐药进化,18个月后耐药病毒数在214例样本中均超过10%的检测标准。结论:LAM治疗在9-18个月获得良好的治疗效果,建议24个月后停药更换临床处方防止耐药病毒株的进化。     

I b 型慢性丙肝患者血清IL-10、IL-12 水平 与HCV-RNA 载量关系的研究*

目的:研究I b型慢性丙肝患者血清IL-10、IL-12水平与HCV-RNA载量的关系,探讨难治性丙肝的免疫发病机制。方法:选取HCV-RNA阳性的I b型慢性丙肝患者48例,依据病毒载量将其分为三个组,低病毒载量组(1.0*103IU/mL1.0*106IU/mL)13例,另选HCV-RNA阴性的慢性HCV感染者15例,健康献血员15例作为对照组;应用ELISA法检测所有研究对象的血清IL-10、IL-12水平。结果:慢性丙肝患者外周血IL-10水平显著高于健康献血员(P=0.003),IL-12水平低于健康献血员(P=0.045);随着HCV-RNA载量的升高,HCV-RNA阳性的I b型慢性患者外周血IL-10水平逐渐升高,组间比较差异具有统计学意义(F=18.65,P=0.000);血清HCV-RNA载量与血清IL-10水平呈正相关(相关系数r=0.71,P=0.000),与血清IL-12水平无相关性(P=0.479)。结论:慢性HCV感染者与患者的细胞免疫均倾向于TH2型;I b型慢性丙肝患者外周血IL-10水平与HCV-RNA病毒载量呈正相关。     

妊娠后期应用替比夫定阻断HBV高载量孕妇母婴传播效果及安全性

目的评价妊娠后期应用替比夫定(LdT)阻断HBV高载量孕妇母婴传播的疗效及安全性。方法选择185例2012年5月1日至2013年4月30日在余姚市人民医院门诊就诊及住院的HBsAg阳性、HBV DNA≥2.0×105 IU/mL、肝功能正常的妊娠妇女,按照患者意愿分为治疗组(127例,自孕28周至产后30d口服LdT 600mg/d)和对照组(58例,未服药)。两组孕妇均在孕26~27周、分娩时及产后30d检测HBV DNA。两组婴儿在出生后6h内、30d、6个月注射乙型肝炎疫苗10μg,同时在婴儿出生后6h内及产后30d注射乙型肝炎免疫球蛋白200IU。在婴儿7~8月龄时检测HBsAg情况。结果治疗组孕妇在分娩时及产后30dHBV DNA滴度明显下降,HBV DNA的阴转率为22.04%,在分娩时HBV DNA水平与对照组相比差异有统计学意义(P=0.001);治疗组婴儿HBsAg阳性率为0.00%,对照组为5.17%(3例感染),两组比较差异有统计学意义(P=0.01)。两组患者在剖宫产率、产后出血率、早产率、新生儿先天畸形率、正常体重儿等方面差异无统计学意义。结论 HBV DNA高载量孕妇在妊娠后期实施抗病毒治疗,能有效抑制孕妇体内HBV DNA水平,降低HBV母婴垂直传播率。     

HBV相关肝纤维化小鼠模型的建立及关键表型深度分析研究

目的 探索HBV感染所致肝纤维化小鼠模型形成条件及表型分析。方法 以18只CBA/CaJ小鼠为研究对象,随机分为对照组(n=4)和实验组(n=14),实验组按照每只2μg的剂量、通过高压水动力方法进行尾静脉注射cccDNA,对照组注射等体积PBS,转染68周后采样。通过qPCR方法检测肝中HBV DNA及HBV cccDNA,ELISA方法检测血清HBsAg、HBeAg,免疫组化检测肝组织中HBsAg、HBcAg,用一代测序法检测HBeAg⁺/HBsAg^-及HBeAg^-/HBsAg⁺组织DNA基因测序,生化分析ALT和AST,HE染色、天狼星染色及Masson染色分析肝组织病理。结果 实验组中,100%的小鼠HBV-DNA拷贝数高于1000 copies/mL;小鼠肝组织cccDNA拷贝数显著高于空白组(P<0.05);42.9%的小鼠血清HBsAg和HBeAg呈阳性;60%的小鼠肝HBsAg呈阳性和26.7%的小鼠肝HBcAg呈阳性;HBeAg^-/HBsAg^+...     

乙肝病毒携带者外周血单核细胞miR-122表达与病毒增殖的关系研究

乙肝病毒携带者外周血单核细胞(Peripheral blood mononuclear cell,PBMC)中微小RNA(microRNA,miR)-122的表达情况及其与病毒增殖的关系,目前鲜有报道。本研究通过检测乙肝病毒携带者PBMC中miR-122的表达情况,探讨其与病毒增殖的关系。选取2017年1月～2018年8月石嘴山中心医院内科门诊的58例乙肝病毒携带者为观察组研究对象,按HBeAg的状态将其分为:HBeAg阳性(+)组31例,HBeAg阴性(-)组27例,同期选择健康体检者32例作为对照组,采用时间分辨荧光免疫分析(Timed-resolved fluoroimmunoassay,TRFIA)定量检测乙肝两对半,采用实时定量PCR法检测血清中HBV DNA拷贝数,采用qRT-PCR法检测PBMC中miR-122表达水平,采用Pearson相关性分析分析乙肝病毒携带者PBMC中miR-122水平与血清HBV DNA拷贝数的关系。结果显示,HBeAg(+)组HBV DNA拷贝数显著高于HBeAg(-)组(P0.05);观察组PBMC中miR-122表达水平显著高于对照组(P0.001);HBeAg(+)组外周血单个核细胞miR-122表达水平显著高于HBeAg(-)组(P0.001);乙肝病毒携带者PBMC中miR-122水平与HBV DNA拷贝数呈正相关(r=0.501,P0.001)。本研究中,与HBeAg(-)乙肝病毒携带者相比,HBeAg(+)乙肝病毒携带者病毒复制活跃,HBV DNA拷贝数升高,miR-122表达上调,提示乙肝病毒携带者PBMC中miR-122表达异常可能与病毒增殖有关。     

CXCL9 Associated with Sustained Virological Response in Chronic Hepatitis B Patients Receiving Peginterferon Alfa-2a Therapy: A Pilot Study

Background and Aims

There is lack of a practical biomarker to predict sustained virological response (SVR) in chronic hepatitis B (CHB) patients undergoing peginterferon alfa-2a (PEG-IFN). The aim of this pilot study was to identify immunological features associated with SVR.

Methods

Consecutive 74 CHB patients receiving 24 weeks (for hepatitis B e antigen (HBeAg)-positive) or 48 weeks (for HBeAg-negative) PEG-IFN, were prospectively enrolled. Serum HBV viral loads, hepatitis B surface antigen (HBsAg), CXCL9, IFN-γ-inducible protein 10 (IP-10), interferon-gamma (IFN-γ) and transforming growth factor beta (TGF-β) were measured at baseline and week 12. SVR was defined as HBeAg seroconversion combined with viral load <2000 IU/mL in HBeAg-positive (n=36), and viral load <2000 IU/mL in HBeAg-negative patients (n=38) at 48 weeks after the end of treatment.

Results

Nineteen patients (25.7%), 7 in HBeAg-positive and 12 in HBeAg-negative, achieved SVR. There were significant declines of HBV DNA, HBsAg, IP-10 and IFN-γ levels at week 12. In multivariate analysis, pre-treatment CXCL9 >80 pg/mL, HBV DNA <2.5 x 10⁷ IU/mL and on-treatment HBV viral load, HBsAg decline >10% at week 12 were predictors of SVR. The performance of CXCL9 in predicting SVR was good in patients with HBV DNA <2.5 x 10⁷ IU/mL, particularly in HBeAg-negative CHB cases (positive predictive value, PPV= 64.3%).

Conclusions

Pre-treatment CXCL9 level has the potential to select CHB patients who can respond to PEG-IFN, especially in HBeAg-negative patients with low viral loads.     

Early Serum HBsAg Kinetics as Predictor of HBsAg Loss in Patients with HBeAg-Negative Chronic Hepatitis B after Treatment with Pegylated Interferonα-2a

Hepatitis B surface antigen (HBsAg) loss is an ideal treatment endpoint for patients with chronic hepatitis B (CHB). We investigated the predictive value of on-treatment HBsAg levels for HBsAg loss in hepatitis B e antigen (HBeAg)-negative CHB patients who received 120-week PEG-IFNα-2a treatment. Serum HBV DNA, HBsAg, and anti-HBs levels were assayed at baseline and every 3 months during the treatment. Of 81 patients, 12 achieved HBsAg loss, 20 achieved HBsAg < 100 IU/mL, and 49 maintained HBsAg≥100 IU/mL. HBsAg loss rate was only 3.7% at 48 weeks, while it reached to 11.1% and 14.8% after treatment of 96 weeks and 120 weeks. The cutoff HBsAg levels at 12 weeks predicting HBsAg loss at 96 weeks and 120 weeks of treatment were 400 IU/mL and 750 IU/mL, with AUC 0.725 and 0.722, positive predictive value (PPV) 29.41% and 30.56%, and negative predictive value (NPV) 93.75% and 97.78%, respectively. The cutoff HBsAg levels at 24 weeks predicting HBsAg loss at 96 weeks and 120 weeks of treatment were 174 IU/mL and 236 IU/mL respectively, with AUC 0.925 and 0.922, PPV 40.0% and 46.15%, and both NPV 100%. The predictive ability of the cutoff HBsAg levels at 24 weeks was better than that at 12 weeks for HBsAg loss at either 96 or 120 weeks (χ²=3.880, P=0.049 and χ²=4.412, P=0.036). These results indicate that extended therapy is critical to HBsAg loss in HBeAg-negative CHB patients during PEG-IFN treatment, and the HBsAg level at 24 weeks can be used to predict HBsAg loss during tailoring PEG-IFN therapy.

     

Dry Blood Spots a Reliable Method for Measurement of Hepatitis B Viral Load in Resource-Limited Settings

MethodsThe performance of DBS in HBV quantification was investigated using a modified commercial test (Abbott RealTime HBV assay). Paired DBS and plasma samples were collected from an HBV positive cohort in Addis Ababa, Ethiopia. DBS were stored at ambient temperature for 4–39 days before shipment to the laboratory.ResultsTwenty-six paired samples were selected covering the total range of quantification, from 2.14 log IU/ml to >7 log IU/ml. HBV was detected in 21 of 21 (100%) DBS from patients with a corresponding plasma viral load above 2.70 log IU/ml. The mean difference between plasma and DBS was 0.59 log IU/ml, and the correlation was strong (R2 = 0.92). In stability studies there was no significant change in DBS viral load after storage at room temperature for up to 12 weeks.ConclusionsThis study suggests that DBS can be a feasible and reliable alternative to plasma for quantification of HBV in resource-limited settings. DBS can expand access to antiviral treatment for patients in low- and middle-income countries.     

Pegylated-Interferon Alpha Therapy for Treatment-Experienced Chronic Hepatitis B Patients

Background

Studies are limited on pegylated interferon (Peg-IFN) therapy for chronic hepatitis B (CHB) patients who failed or relapsed on previous antiviral therapy.

Objectives

We aimed to investigate the effect of Peg-IFN therapy in treatment-experienced CHB patients.

Study Design

A total of 57 treatment-experienced CHB patients at two medical centers were enrolled. All of the patients were treated with Peg-IFN α-2a at 180 μg weekly for 24 or 48 weeks. The hepatitis B serological markers and viral loads were tested every 3 months until 1 year after stopping Peg-IFN therapy. The endpoints were HBV DNA <2000IU/mL, hepatitis B e antigen (HBeAg) seroconversion, and a hepatitis B surface antigen (HBsAg) loss at 12 months post-treatment.

Results

In HBeAg-positive patients, 25.0%, 29.2%, and 12.5% of the patients achieved HBeAg seroconversion, HBV DNA <2000 IU/mL and a combined response, respectively, at 12 months post-treatment. Prior IFN therapy, a high baseline ALT level, a low creatinine level, undetectable HBV DNA at 12 weeks and a decline in HBV DNA >2 log₁₀ IU/mL at 12 weeks of therapy were factors associated with treatment response. In HBeAg-negative patients, 9.1%, 15.2%, and 6.1% of the patients achieved undetectable HBV DNA, HBV DNA <2000 IU/mL, and an HBsAg loss, respectively, at 12 months post-treatment. No factor was significantly associated with the treatment response in the HBeAg-negative patients. The median HBsAg level declined from 3.4 to 2.6 log₁₀ IU/mL in all the patients, and the 5-year cumulative rate of the HBsAg loss was 9.8% in the HBeAg-negative patients. Overall, none of the patients prematurely discontinued the Peg-IFN therapy.

Conclusions

Peg-IFN re-treatment is effective for a proportion of HBeAg-positive treatment-experienced patients; it has limited efficacy for HBeAg-negative treatment-experienced patients. Peg-IFN might facilitate HBsAg loss in HBeAg-negative treatment-experienced patients.     

Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA

Quantitative real-time PCR (qPCR) has been widely implemented for clinical hepatitis B viral load testing, but a lack of standardization and relatively poor precision hinder its usefulness. Droplet digital PCR (ddPCR) is a promising tool that offers high precision and direct quantification. In this study, we compared the ddPCR QX100 platform by Bio-Rad with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad, USA) to detect serial plasmid DNA dilutions of known concentrations as well as HBV DNA extracted from patient serum samples. Both methods showed a high degree of linearity and quantitative correlation. However, ddPCR assays generated more reproducible results and detected lower copy numbers than qPCR assays. Patient sample quantifications by ddPCR and qPCR were highly agreeable based on the Bland–Altman analysis. Collectively, our findings demonstrate that ddPCR offers improved analytical sensitivity and specificity for HBV measurements and is suitable for clinical HBV detection.     

Levels of hepatitis B virus replicative intermediate in serum samples of chronic hepatitis B patients

Hepatitis B virus (HBV) cccDNA levels is an absolute marker of HBV replication in the liver of HBV infected patients. This study aimed to quantify the HBV cccDNA levels in sera and liver tissue samples of treatment naïve patients with chronic hepatitis B. Eighty one chronic hepatitis B (CHB) treatment naïve patients were enrolled from January 2009 to June 2011. Total HBV DNA and HBV cccDNA levels were quantified using sensitive real time PCR assay. The mean age of recruited patients was 34 ± 11.5 years. Fifty four (66.7 %) patients were HBeAg negative. Liver tissue samples were available from 2 HBeAg positive and 21 HBeAg negative CHB patients. The amount of total intrahepatic HBV DNA ranged from 0.09 to 1508.92 copies/cell. The median intrahepatic HBV cccDNA was 0.31 and 0.20 copies/cell in HBeAg positive and HBeAg negative cases, respectively. Serum HBV cccDNA was detectable in 85.2 % HBeAg positive and 48.1 % HBeAg negative CHB patients. Median serum HBV cccDNA was 46,000 and 26,350 copies/mL in HBeAg positive and HBeAg negative subjects, respectively. There was a significant positive correlation between the levels of intrahepatic total HBV DNA and intrahepatic HBV cccDNA (r = 0.533, p = 0.009). A positive correlation was also seen between serum HBV cccDNA levels and serum HBV DNA levels (r = 0.871, p < 0.001). It was concluded that serum HBV cccDNA could be detectable in higher proportion of HBeAg positive patients compared to HBeAg negative patients. Moreover, the median level of serum HBV cccDNA was significantly higher in HBeAg positive patients in contrast to HBeAg negative subjects.     

A Comparison of Entecavir and Lamivudine for the Prophylaxis of Hepatitis B Virus Reactivation in Solid Tumor Patients Undergoing Systemic Cytotoxic Chemotherapy

Background

Nucleos(t)ide analogues reduce the incidence of hepatitis B virus (HBV) reactivation in cancer patients undergoing systemic cytotoxic chemotherapy but the experience of solid tumors remains limited. Aims. The aim of this study was to compare the efficacy of entecavir and lamivudine in the prophylaxis of HBV reactivation in solid tumor patients undergoing systemic cytotoxic chemotherapy.

Methods

HBsAg seropositive patients undergoing systemic cytotoxic chemotherapy for solid tumors with prophylactic entecavir and lamivudine between January 2006 and June 2013 were retrospectively investigated. The incidence of HBV reactivation and outcome of the patients were analyzed. The risk factors of HBV reactivation were examined.

Results

A total of 213 patients (entecavir group, 70 patients; lamivudine group, 143 patients) were evaluated. Less incidence of HBV reactivation was noticed in entecavir group than in lamivudine group (0% vs. 7.0%, P = 0.02). No HBV reactivation was noticed in the patients with a baseline HBV DNA level < 2000 IU/mL. A baseline HBV DNA level ≥ 2000 IU/mL, HBeAg, and lamivudine were significantly associated with HBV reactivation. Subgroup analysis of the patients with a baseline HBV DNA level ≥ 2000 IU/mL found that lamivudine was significantly associated with HBV reactivation. Most of the reactivation events were properly managed by using tenofovir disoproxil fumarate. The incidence of hepatitis during chemotherapy and disruption of chemotherapy was similar between patients using entecavir and lamivudine with a baseline HBV DNA level ≥ or < 2000 IU/mL.

Conclusions

A baseline HBV DNA level ≥ 2000 IU/mL, HBeAg, and lamivudine were the risk factors of HBV reactivation during systemic cytotoxic chemotherapy in solid tumor patients. Entecavir was superior to lamivudine in terms of less incidence of reactivation in the patients with a baseline HBV DNA level ≥ 2000 IU/mL. Both agents were equally efficacious in the patients with HBV DNA levels < 2000 IU/mL.     

Dried Blood Spot Sampling for Hepatitis B Virus Serology and Molecular Testing

Background & Aims

Dried blood spots (DBS) on filter paper have been successfully used to diagnose and monitor several infectious diseases. The aim was to investigate the performance of DBS in hepatitis B virus (HBV) diagnosis using commercial tests in comparison to standard methods.

Methods

Paired DBS and plasma samples were collected from 200 patients: 100 patients with HBsAg negative status and 100 patients with HBsAg positive status. In the latter patient, HBeAg reactivity was tested. Ten samples of anti-HBs were collected from people vaccinated against HBV. We also studied 50 patients with positive HBV DNA viral load in plasma and 10 HBV DNA negative patients. HBV genotypes and gene polymerase mutations were determined in 10 randomly selected HBV-infected patients. The DBS sample consisted of 50 µL of whole blood, i.e. a 12-mm paper card.

Results

The sensitivity thresholds of HBsAg and anti-HBs antibody were 0.30±0.08 IU/mL and 18.11±6.05 IU/mL, respectively, for DBS with 98% sensitivity and 100% specificity. Sensitivity was 98% and specificity 100% for the detection of HBV DNA on a blotter, considering an HBV DNA threshold of 914.1±157.8 IU/ml. Ten patients had an HBeAg positive status in plasma, all were detected positive using DBS. HBV genotyping and mutation detection were successfully performed on DBS, with full concordance between the 10 paired DBS and plasma samples.

Conclusion

This study shows DBS is a reliable alternative to plasma specimens for quantifying and detecting HBsAg, anti-HBs, HBeAg and genotyping. DBS may increase the opportunities for HBV testing and treatment follow-up in hard-to-reach individuals.     

Comparison of the Efficacy of Entecavir and Tenofovir in Nucleos(T)ide Analogue-Experienced Chronic Hepatitis B Patients

The efficacy of entecavir (ETV) and tenofovir (TDF) for the treatment of nucleos(t)ide analogue (NA)-experienced chronic hepatitis B (CHB) patients has been little studied. Here, we compare the efficacy of both ETV and TDF in NA-experienced CHB patients without detectable genotypic resistance. This retrospective cohort study included consecutive NA-experienced patients who had neither current nor previous genotypic resistance and had received ETV or TDF for at least 6 months. Overall, 202 patients (146 patients in the ETV group and 56 in the TDF group) were analyzed. The cumulative probabilities of complete virologic suppression (CVS) at month 12 were 76.1% in the ETV group and 95.0% in the TDF group (P<0.001), respectively. The TDF-treated group achieved CVS more rapidly than the ETV group for both Hepatitis B e antigen (HBeAg)-negative and -positive patients (P = 0.006 and < 0.001, respectively), and for those with both low (< 2,000 IU/mL) and high (≥ 2,000 IU/mL) HBV DNA levels (P = 0.01 and 0.002, respectively). TDF group had an increased probability of achieving CVS (hazard ratio, 2.242; 95% confidence interval, 1.587–3.165; P = 0.001), after adjustment for HBV DNA level, the presence of HBeAg, and a history of CVS during prior treatment. During the treatment period, 23 patients (15.8%) in the ETV group developed virologic breakthrough, compared to none in the TDF group. The cumulative probabilities of developing virologic breakthrough and ETV-resistance at month 24 were 9.7% and 5.3%, respectively. In conclusion, TDF is preferable to ETV for achieving CVS in NA-experienced CHB patients without genotypic resistance.